Signaling by the ERK-MAP kinases and RSK family of protein kinases plays a significant role in the regulation of processes controlling cell growth and proliferation, cell migration and invasion, cell survival, and differentiation. Thus, when improperly regulated, ERK/RSK signaling also contribute to the neoplastic state. Indeed, several oncogenes and their cellular homologs have been found to directly participate in the regulation of ERK/RSK signaling and well over 50% of human cancers have this pathway activated. Therefore, it is critical to thoroughly define ERK/RSK regulatory mechanisms and their downstream effectors. The original goals during the last funding period (10 year MERIT award) were to characterize RSK regulation by ERK and other inputs, identify RSK interacting proteins and identify new RSK targets. The ultimate goal was to define the molecular and biochemical basis of ERK and RSK activation, downstream signaling and how these events regulate various biological processes including transcriptional and translational control, cell migration and cell survival. These goals evolved to include characterization of ERK signaling via distinct interaction motifs and the biological consequences. We have made several significant discoveries that continue to further our understanding of the regulation and functional consequences of ERK/RSK signaling in normal health and when improperly regulated, diseases such as cancer. Our previous work and that of others sets the foundation for the current proposal. From our innovative approaches and proposed research, we will define the ERK/RSK phosphoproteome and proteome using a unique ERK signaling system we have developed and a reductive dimethylation/mass spectrometry approach (aim 1). The initial data have already been obtained after acute EGF stimulation and this is to be expanded so that we will define the changing proteome/phosphoproteome with a time course analysis to inform us as to how cells make specific cell fate decisions.
Aim 1 also includes gene expression analysis during this time course to complement the mass spectrometry, and has already revealed genes specific to ERK/RSK-regulated cell proliferation vs. genes specific to ERK-regulated epithelial to mesenchymal transition.
Aim 2 will use data from the screen to initiate our efforts to define cross-talk pathways and negative feedback loops regulated by RSK that contribute to signaling and cell fate decisions. Phosphoproteomic data and published results will also be used to characterize signaling linking RSK to cell migration and survival via regulation of the actomyosin cytoskeleton (aim 3). Through these aims, we will identify and characterize new upstream regulators and downstream effectors of ERK/RSK, further explaining how improper regulation of the Ras pathway contributes to a variety of human diseases. Furthermore, our research will result in the identification of new biomarkers and potential therapeutic targets needed for detection and intervention in diseases resulting from improper ERK/RSK signaling.

Public Health Relevance

With the combination of hypothesis-driven and hypothesis-generating approaches described in this proposal, and the vast amount of data to be generated, we will establish a new level of understanding of Ras oncoprotein and growth factor driven ERK/RSK signal transduction and its links to several of the major hallmarks of cancer, including improper regulation of cell proliferation, cell survival, and cell migration/invasion. With the described aims, we have placed ourselves in the unique position to uncover and understand at a biochemical and molecular level new regulatory processes, new biomarkers and new potential targets for drug discovery, that are needed for personalized therapeutic intervention in many diseases, such as cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA046595-27A1
Application #
8576568
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Watson, Joanna M
Project Start
1988-02-08
Project End
2018-04-30
Budget Start
2013-07-01
Budget End
2014-04-30
Support Year
27
Fiscal Year
2013
Total Cost
$689,580
Indirect Cost
$278,607
Name
Harvard University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
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Shin, Sejeong; Buel, Gwen R; Wolgamott, Laura et al. (2015) ERK2 Mediates Metabolic Stress Response to Regulate Cell Fate. Mol Cell 59:382-98
Mendoza, Michelle C; Vilela, Marco; Juarez, Jesus E et al. (2015) ERK reinforces actin polymerization to power persistent edge protrusion during motility. Sci Signal 8:ra47
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Galan, Jacob A; Geraghty, Kathryn M; Lavoie, Geneviève et al. (2014) Phosphoproteomic analysis identifies the tumor suppressor PDCD4 as a RSK substrate negatively regulated by 14-3-3. Proc Natl Acad Sci U S A 111:E2918-27
Yi, Tingfang; Zhai, Bo; Yu, Yonghao et al. (2014) Quantitative phosphoproteomic analysis reveals system-wide signaling pathways downstream of SDF-1/CXCR4 in breast cancer stem cells. Proc Natl Acad Sci U S A 111:E2182-90
Gu, Xiaoxiao; Yu, Jane J; Ilter, Didem et al. (2013) Integration of mTOR and estrogen-ERK2 signaling in lymphangioleiomyomatosis pathogenesis. Proc Natl Acad Sci U S A 110:14960-5
Kim, Sang Gyun; Buel, Gwen R; Blenis, John (2013) Nutrient regulation of the mTOR complex 1 signaling pathway. Mol Cells 35:463-73
Er, Ekrem Emrah; Mendoza, Michelle C; Mackey, Ashley M et al. (2013) AKT facilitates EGFR trafficking and degradation by phosphorylating and activating PIKfyve. Sci Signal 6:ra45

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